Dynamic remodeling of subcellular chemical gradients using a multi-directional flow device
Dynamic remodeling of subcellular chemical gradients using a multi-directional flow device
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DOI：
10.1039/c004627b
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发表时间：
2010-01-01
期刊：
影响因子：
6.1
通讯作者：
Shear, Jason B.
中科院分区：
文献类型：
--
作者：
Moorjani, Samira;Nielson, Rex;Shear, Jason B.
阐明外部化学信号调节极化细胞行为的机制需要能够快速重塑亚细胞分辨率化学景观的工具。在这里，我们描述了一种方法，用于创建陡峭的微观梯度的细胞效应器在任何所需的位置，在文化中，可以重新定位迅速评估动态响应。在这种方法中，在支持细胞粘附的膜中烧蚀微米孔，从而允许来自底层储器的定量试剂作为尖锐流进入细胞培养流动室，所述尖锐流通过使用成对的源和排出口的系统来指定流动方向而指向亚细胞靶。该工具大大扩展了与培养细胞的化学相互作用的能力，使得能够通过精确放置和重新定向在给药流的边界处形成的肽梯度来研究趋化性。这些研究表明，中性粒细胞前体细胞可以revert和重定向其迁移路径使用形态学反应，依赖于亚细胞定位的趋化梯度。
Elucidation of the mechanisms by which external chemical cues regulate polarized cellular behaviors requires tools that can rapidly recast chemical landscapes with subcellular resolution. Here, we describe an approach for creating steep microscopic gradients of cellular effectors at any desired position in culture that can be reoriented rapidly to evaluate dynamic responses. In this approach, micrometre pores are ablated in a membrane that supports cell adherence, allowing dosing reagent from an underlying reservoir to enter the cell-culture flow chamber as sharp streams that are directed at subcellular targets by using a system of paired sources and drains to specify flow direction. This tool substantially extends capabilities for chemical interaction with cultured cells, enabling investigations of chemotaxis via precise placement and reorientation of peptide gradients formed at the boundaries of dosing streams. These studies demonstrate that neutrophil precursor cells can repolarize and redirect their migration paths using morphological responses that depend on the subcellular localization of chemoattractant gradients.